Influence of Pulsed Electric Fields and Mitochondria-Cytoskeleton Interactions on Cell Respiration.
Ontology highlight
ABSTRACT: Pulsed electric fields with microsecond pulse width (?sPEFs) are used clinically; namely, irreversible electroporation/Nanoknife is used for soft tissue tumor ablation. The ?sPEF pulse parameters used in irreversible electroporation (0.5-1 kV/cm, 80-100 pulses, ?100 ?s each, 1 Hz frequency) may cause an internal field to develop within the cell because of the disruption of the outer cell membrane and subsequent penetration of the electric field. An internal field may disrupt voltage-sensitive mitochondria, although the research literature has been relatively unclear regarding whether such disruptions occur with ?sPEFs. This investigation reports the influence of clinically used ?sPEF parameters on mitochondrial respiration in live cells. Using a high-throughput Agilent Seahorse machine, it was observed that ?sPEF exposure comprising 80 pulses with amplitudes of 600 or 700 V/cm did not alter mitochondrial respiration in 4T1 cells measured after overnight postexposure recovery. To record alterations in mitochondrial function immediately after ?sPEF exposure, high-resolution respirometry was used to measure the electron transport chain state via responses to glutamate-malate and ADP and mitochondrial membrane potential via response to carbonyl cyanide-p-trifluoromethoxyphenylhydrazone. In addition to measuring immediate mitochondrial responses to ?sPEF exposure, measurements were also made on cells permeabilized using digitonin and those with compromised cytoskeleton due to actin depolymerization via treatment with the drug latrunculin B. The former treatment was used as a control to tease out the effects of plasma membrane permeabilization, whereas the latter was used to investigate indirect effects on the mitochondria that may occur if ?sPEFs impact the cytoskeleton on which the mitochondria are anchored. Based on the results, it was concluded that within the pulse parameters tested, ?sPEFs alone do not hinder mitochondrial physiology but can be used to impact the mitochondria upon compromising the actin. Mitochondrial susceptibility to ?sPEF after actin depolymerization provides, to our knowledge, a novel avenue for cancer therapeutics.
SUBMITTER: Goswami I
PROVIDER: S-EPMC6026445 | biostudies-literature | 2018 Jun
REPOSITORIES: biostudies-literature
ACCESS DATA